摘要
目的探讨Six1对人支气管上皮细胞(16HBE)间质转化(EMT)及纤维化的影响。方法体外培养16HBE细胞株并分成3组,实验组转染Six1(NM_005982)-GV146质粒,阴性对照组转染GV146对照质粒,空白对照组仅给予LipofectamineTM2000转染试剂。转染48 h后用倒置显微镜观察各组细胞形态的变化;采用RT-PCR及Western blot技术检测Six1、E-钙黏蛋白、α肌动蛋白、波形蛋白、纤维粘连蛋白、角蛋白的mRNA和蛋白表达。结果与阴性对照组及空白对照组比较,实验组细胞间隙增大、形态发生梭形改变;Six1、α肌动蛋白、波形蛋白、纤维粘连蛋白的mRNA及蛋白表达增高,差异有显著性(F=11.40~881.42,P<0.05);E-钙黏蛋白、角蛋白mRNA及蛋白表达明显降低,差异有显著性(F=10.71~20.37,P<0.05)。阴性对照组与空白对照组以上各mRNA及蛋白的表达比较差异均无统计学意义(P>0.05)。结论Six1过表达可使16HBE细胞发生EMT和纤维化改变。
Objective To investigate the effect of Six1 on epithelial-mesenchymal transition(EMT)and fibrosis in human bronchial epithelial cells(16HBE).Methods 16HBE cells were cultured in vitro and then they were divided into experimental group transfected with Six1(NM_005982)-GV146 plasmid,negative control group transfected with GV146 control plasmid,and blank control group treated with the LipofectamineTM 2000 transfection reagent.At 48 hours after transfection,an inverted microscope was used to observe the morphological changes of cells,and RT-PCR and Western blot were used to measure the mRNA and protein expression of Six1,E-cadherin,α-smooth muscle actin(α-SMA),vimentin,fibronectin,and keratin.ResultsCompared with the negative control group and the blank group,the experimental group showed a significant increase in intercellular space and had a spindle shape,as well as significant increases in the mRNA and protein expression of Six1,α-SMA,vimentin,and fibronectin(F=11.40-881.42,P<0.05)and significant reductions in the mRNA and protein expression of E-cadherin and keratin(F=10.71-20.37,P<0.05).There were no significant differences in mRNA and protein expression between the negative control group and the blank control group(P>0.05).Conclusion Six1 overexpression may induce EMT and fibrosis of 16HBE cells.
作者
王文鑫
杨召川
曲政海
王冲
徐雷
张梦雪
WANG Wenxin;YANG Zhaochuan;QU Zhenghai;WANG Chong;XU Lei;ZHANG Mengxue(Depart-ment of Pediatrics, The Affiliated Hospital of Qingdao University, Qingdao 266003, China)
出处
《青岛大学学报（医学版）》
CAS
2020年第4期399-403,共5页
Journal of Qingdao University(Medical Sciences)
基金
国家自然科学基金青年科学基金项目(81700029)
山东省自然科学基金项目(ZR2017MH017)
青岛市民生科技计划项目(19-6-32-nsh)。
关键词
Six1
上皮-间质转化
支气管
上皮细胞
气道重塑
哮喘
Six1
epithelial-mesenchymal transition
bronchi
epithelial cells
airway remodeling
asthma
